Search PubMed⌕ Search

Biomedical subjects

J Seelig

Publications and source records attributed to J Seelig.

At least 37 records · Page 2Linked to original sources
PubMed1999

New drugs for the Na+/H+ exchanger. Influence of Na+ concentration and determination of inhibition constants with a microphysiometer.

The NHE-1 isoform of the Na+/H+ exchanger is excessively activated in cardiac cells during ischemia. Hence NHE-1 specific inhibitors are being developed since they could be of beneficial influence under conditions of cardiac ischemia and reperfusion. In this study, the Cytosensortrade mark microphysiometer was used to measure the potency of four new drug molecules, i.e., EMD 84021, EMD 94309, EMD 96785 and HOE 642 which are inhibitors of the isoform 1 of the Na+/H+ exchanger. The experiments were performed with Chinese hamster ovary cells (CHO K1) which are enriched in the NHE-1 isoform of the Na+/H+ antiporter. The Na+/H+ exchanger was stimulated with NaCl and the rate of extracellular acidification was quantified with the Cytosensor. The proton exchange rate was measured as a function of the NaCl concentration in the range of 10-138 mm NaCl stimulation. The proton exchange rate followed Michaelis-Menten kinetics with a KM = 30 +/- 4 mm for Na+. Addition of either one of the four inhibitors decreased the acidification rate. The IC50 values of the four compounds could be determined as 23 +/- 7 nm for EMD 84021, 5 +/- 1 nm for EMD 94309, 9 +/- 2 nm for EMD 96785 and 8 +/- 2 nm for HOE 642 at 138 mm NaCl, in good agreement with more elaborate biological assays. The IC50 values increased with the NaCl concentration indicating competitive binding of the inhibitor. The microphysiometer approach is a fast and simple method to measure the activity of the Na+/H+ antiporter and allows a quantitative kinetic analysis of the proton excretion rate.

Animals↗
PubMed1999

Metabolic changes in the brain of patients with anorexia and bulimia nervosa as detected by proton magnetic resonance spectroscopy.

OBJECTIVE: To investigate the brain of patients with anorexia and bulimia nervosa by localized proton magnetic resonance spectroscopy (1H-MRS) and to look for metabolic alterations. METHOD: Twenty patients with anorexia and bulimia nervosa were investigated by magnetic resonance imaging (MRI) and 1H-MRS in three regions of the brain. Age and sex-matched healthy subjects were investigated as controls. RESULTS: 1H-MRS revealed metabolic changes, such as a significant decrease of both myo-inositol and lipid compounds within the frontal white matter. The concentration of these compounds was further reduced with decreasing body mass index. Reduced lipid signals were also found in the occipital gray matter. In the cerebellum, the concentration of all metabolites including water, except lipids, was increased. DISCUSSION: The metabolic changes found in this study seem to be a consequence of nutritional deficiency. It has to be further investigated whether these findings have any relevance for brain function. 1H-MRS might serve as a valuable investigative tool to observe eating disorders as anorexia and bulimia nervosa and to follow the success of therapy.

Adolescent↗
PubMed1999

19F-MRI of perfluorononane as a novel contrast modality for gastrointestinal imaging.

19F-Magnetic resonance imaging in conjunction with perfluorononane provides a new modality for gastrointestinal (GI) imaging as is demonstrated here with an animal model. Perfluorononane was found to be an ideal oral contrast agent since it is biologically inert, immiscible with water, and since it has a low viscosity and surface tension. Furthermore, its high fluorine content, together with the high sensitivity of 19F-MRI, allowed highly selective MR images of the GI tract of mice to be acquired. Due to the lack of 19F background signals, the contrast of the GI tract was only limited by the signal-to-noise ratio of the 19F-MR images. 19F-RARE images of 1-mm slices with an in-plane resolution of 0.23 x 0.23 mm2 were obtained from the GI tract after oral perfluorononane administration. The passage of perfluorononane through the entire GI tract was monitored by repetitive MR measurements with a maximal time resolution of 38 s. The three-dimensional surfaces of the GI tract were reconstructed and superimposed on corresponding 1H-MR images, which provided complementary anatomical information.

Administration, Oral↗
PubMed1998

Interaction of the neuronal marker dye FM1-43 with lipid membranes. Thermodynamics and lipid ordering.

The fluorescent dye FM1-43 labels nerve terminals in an activity-dependent fashion and has been found increasingly useful in exploring the exo- and endocytosis of synaptic vesicles and other cells by fluorescence methods. The dye distributes between the aqueous phase and the lipid membrane but the physical-chemical parameters characterizing the adsorption/partition equilibrium have not yet been determined. Fluorescence spectroscopy alone is not sufficient for a detailed elucidation of the adsorption mechanism since the method can be applied only in a rather narrow low-concentration window. In addition to fluorescence spectroscopy, we have therefore employed high sensitivity isothermal titration calorimetry (ITC) and deuterium magnetic resonance (2H-NMR). ITC allows the measurement of the adsorption isotherm up to 100 microM dye concentration whereas 2H-NMR provides information on the location of the dye with respect to the plane of the membrane. Dye adsorption/partition isotherms were measured for neutral and negatively-charged phospholipid vesicles. A non-linear dependence between the extent of adsorption and the free dye concentration was observed. Though the adsorption was mainly driven by the insertion of the non-polar part of the dye into the hydrophobic membrane interior, the adsorption equilibrium was further modulated by an electrostatic attraction/repulsion interaction of the cationic dye (z=+2) with the membrane surface. The Gouy-Chapman theory was employed to separate electrostatic and hydrophobic effects. After correcting for electrostatic effects, the dye-membrane interaction could be described by a simple partition equilibrium (Xb=Kcdye) with a partition constant of 103-104 M-1, a partition enthalpy of DeltaH=-2.0 kcal/mol and a free energy of binding of DeltaG=-7.8 kcal/mol. The insertion of FM1-43 into lipid membranes at room temperature is thus an entropy-driven reaction following the classical hydrophobic effect. Deuterium nuclear magnetic resonance provided insight into the structural changes of the lipid bilayer induced by the insertion of FM1-43. The dye disturbed the packing of the fatty acyl chains and decreased the fatty acyl chain order. FM1-43 also induced a conformational change in the phosphocholine headgroup. The -P-N+ dipole was parallel to the membrane surface in the absence of dye and was rotated with its positive end towards the water phase upon dye insertion. The extent of rotation was, however, much smaller than that induced by other cationic molecules of similar charge, suggesting an alignment of FM1-43 such that the POPC phosphate group is sandwiched by the two quaternary FM1-43 ammonium groups. In such an arrangement the two cationic charges counteract each other in a rotation of the -P-N+ dipole.

Calorimetry↗
PubMed1998

Proton induced vesicle fusion and the isothermal lalpha-->HII phase transition of lipid bilayers: a 31P-NMR and titration calorimetry study.

The proton-induced isothermal fusion of unilamellar lipid vesicles (Duzgunes et al., Biochemistry 24 (1985) 3091-3098) is compared with the lamellar (Lalpha)-->hexagonal (HII) phase transition of multilamellar lipid dispersions. Both lipid systems are composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and oleic acid (OA) at a 7:3 molar ratio. Using solid-state phosphorus-31 nuclear magnetic resonance (31P-NMR) it is demonstrated that the multilamellar lipid dispersions are in the bilayer state at physiological pH and undergo a Lalpha-->HII phase transition between pH 6.3 and 5.7. This phase transition can also be induced at constant pH by increasing the temperature. The midpoint of the temperature-induced Lalpha-->HII transition is Th=56 degrees C (at pH 7.4) and the corresponding transition enthalpy is DeltaH=0. 7+/-0.1 kcal/mol as determined with differential scanning calorimetry. Both the proton-induced and the temperature-induced phase transition can be completely inhibited by addition of 30 mol% of 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (LPC). In a second set of experiments unilamellar vesicles are prepared either by sonication or by extrusion through polycarbonate filters at pH 7. 4 and are titrated into buffer at pH 5.7. The proton-induced fusion of the lipid vesicles is monitored with isothermal titration calorimetry, light scattering and fluorescence spectroscopy. The fusion reaction is characterized by an endothermic enthalpy of DeltaH=0.5+/-0.2 kcal/mol (at 28 degrees C). The fusion enthalpy is independent of the vesicle diameter and is only slightly reduced by an increase in temperature to 50 degrees C. Vesicle fusion is accompanied by an increase in light scattering, indicating the formation of larger lipid structures. The transition from unilamellar vesicles to fused lipid structures occurs in the same narrow pH range of 6.3-5.7 as observed for the Lalpha-->HII transition of multilamellar dispersions. Vesicle fusion can be inhibited with 30% LPC. The virtually identical set of parameters found for the Lalpha-->HII phase transition and the vesicle fusion reaction suggests that vesicle fusion also entails a Lalpha-->HII phase transition.

Calorimetry↗
PubMed1998

Magainin 2 amide interaction with lipid membranes: calorimetric detection of peptide binding and pore formation.

The interaction of the antibiotic magainin 2 amide (M2a) with lipid bilayers was investigated with high-sensitivity titration calorimetry. The enthalpy of transfer of the cationic M2a to negatively charged small unilamellar vesicles composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG) (75:25, mol/mol) was measured as delta H = -17.0 +/- 1 kcal/mol of peptide. The adsorption isotherm was determined by injecting lipid vesicles into peptide solutions at low peptide concentrations (cPo < 7 microM). The apparent partition coefficient was Kapp approximately 1.2 x 10(4) M-1 at a peptide equilibrium concentration of 1 microM but decreased with increasing peptide concentration. The hydrophobic partitioning of M2a into the lipid membrane is modulated by electrostatic effects that arise from the attraction of the positively charged peptide to the negatively charged membrane. Using the Gouy-Chapman theory to correct for electrostatic attraction, the experimental binding isotherms can be explained with an intrinsic (hydrophobic) partition coefficient of K = 55 +/- 5 M-1 and an effective peptide charge of z = 3.7-3.8. The free energy of binding is delta G = -4.8 kcal/mol. At peptide concentrations cPo > approximately 7 microM, a second effect comes into play, and the titration enthalpies can no longer be explained exclusively by peptide partitioning. The first few injections produce enthalpies of reaction which are distinctly smaller than expected from a pure partition equilibrium, followed by a series of injections with reaction heats larger than expected. After subtracting the enthalpic contribution due to partitioning, the residual enthalpies are endothermic for the first few injections, and exothermic for the consecutive steps. Furthermore, the endothermic excess heat is compensated exactly by the exothermic excess heat; i.e., the excess heat consumed in the first part of the titration experiment is returned during the second part. Endothermic excess enthalpies are observed for total molar peptide-to-lipid ratios of P/L > approximately 3.0%, whereas exothermic excess heats were seen for 0.7% < P/L < 3.0%. Below P/L < approximately 0.7%, the binding follows the partition equilibrium. Based on earlier spectroscopic evidence, it is suggested that magainin 2 amide binds to the lipid membrane and forms pores at high peptide-to-lipid ratio, this process being characterized by an endothermic reaction enthalpy. Pore formation is reversed with increasing lipid concentration, and the peptide pores disintegrate. The limiting peptide-to-lipid ratio deduced from titration calorimetry for M2a pore formation is in excellent agreement with spectroscopic methods. The enthalpy of pore formation amounts to delta H = +6.2 +/- 1.6 kcal/mol peptide or delta H approximately 25-45 kcal/mol pore if the pore is comprised of 4-7 peptide molecules.

Amino Acid Sequence↗
PubMed1998

29Si imaging of silicone breast implants and intraocular silicone oil.

Silicon-29 (29Si) imaging was investigated as a potential imaging modality for monitoring silicone prostheses in humans. The 29Si relaxation times of several silicone gels were measured and found to average T1 = 21.2 +/- 1.5 s and T2 = 207 +/- 40 ms, with no significant difference between virgin and explanted gels. A single-shot half-Fourier rapid acquisition with relaxation enhancement (RARE) and a refocused gradient-echo sequence were used for acquiring 29Si images with 5 x 5 mm2 resolution and no slice selection. Three volunteers with silicone-gel-filled breast implants and one subject with an intraocular silicone oil injection were thus examined in a total acquisition time of 10-15 min per image. On all 29Si images, the shape of the silicone object was well depicted. Although at present, conventional proton images are superior in resolution and signal-to-noise ratio, 29Si imaging has the advantage of optimal specificity, since only the silicone itself is visible.

Biocompatible Materials↗
PubMed1998

Tonotopic organization of the human auditory cortex as detected by BOLD-FMRI.

Functional magnetic resonance imaging is a noninvasive and nonradioactive method for the detection of focal brain activity. In the present study the auditory cortex was investigated in nine normal subjects who were binaurally stimulated using pulsed sine tones of 500 Hz and 4000 Hz. The BOLD (blood oxygenation level dependent) signal change coincided with the stimulation paradigm and was detected in the plane of the superior temporal gyrus. The comparison of the spatial distribution of activated areas revealed a different behavior for the two frequencies. The present findings underline the existence of a frequency specific organization in the medio-lateral, fronto-occipital and cranio-caudal extension in both hemispheres of the auditory cortex in human. The activated areas for the high tone were found more frontally and medially orientated than the low tone stimulated areas. Furthermore, a slight cranio-caudal shift was observed for the higher frequency, more pronounced in the right than in the left temporal lobe. Finally, for most of the subjects investigated the BOLD activation area of the 500 Hz sine tone was larger than that of the 4000 Hz stimulation. Both frequencies showed a lateralization of signal response to the left temporal lobe.

Acoustic Stimulation↗
PubMed1998

[Visualization of central auditory processes with functional magnetic resonance tomography].

BACKGROUND: Central auditory processes can be visualized using functional MRI in a non-invasive manner and at high spatial resolution. Acoustic stimulation leads to an increase of blood flow of activated areas in the plane of the superior temporal gyrus. Radiologically, this may be visualized based on the long T2-relaxation time of oxyhemoglobin. PATIENTS: Ten normal-hearing subjects with ages between 28 and 38 years took part in the investigations. They received binaural, monaural right, and monaural left stimulation with pulsed sine tones of 1000 Hz at a pulse rate of 6 Hz and a sound pressure level of 100 dB SPL. Tonotopic organization of the auditory cortex was visualized using stimulation by pulsed sine tones of 500 Hz and 4000 Hz. RESULTS: Following monaural acoustic stimulation, increased activity of the contralateral auditory cortex could be demonstrated in 9 subjects. In one subject, bilateral activity was noted. Concerning the tonotopic organization of the auditory cortex, we could show that the higher frequencies were localized more medially and anteriorly; the lower frequencies were localized more laterally and posteriorly in the superior temporal gyrus. However, considerable overlap was noted. CONCLUSIONS: The overlap of the different frequencies could explain the controversial discussion of the tonotopic organization of the auditory cortex. The results of the monaural acoustic stimulation show clearly the predominant signal increase of contralateral areas in the primary auditory cortex. These results confirm the opinion of the current textbooks that the fiber of the auditory pathways mostly cross. Further investigations using functional MRI are necessary for better understanding of physiological and pathophysiological central-auditory processes.

Acoustic Stimulation↗
PubMed1998

Auditory cortical responses in hearing subjects and unilateral deaf patients as detected by functional magnetic resonance imaging.

Functional magnetic resonance imaging is a non-invasive method for the detection of focal brain activity at high spatial resolution. Acoustic stimulation leads to a blood oxygenation level dependent signal change in the plane of the superior temporal gyrus. The dependence of this response in the auditory cortex on binaural, monaural left and monaural right acoustic stimulation for 10 healthy subjects and five monaural deaf patients is described. Acoustic stimulation consists of 1000 Hz pulsed sine tones at a pulse rate of 6 Hz and a sound pressure level of 95 dB. For monaural stimulation, normal-hearing subjects revealed a strong lateralization of cortical response towards the contralateral hemisphere. The lateralization ratios between left and right hemispheric response areas were 3.4-5.2 for monaural stimulation and nearly balanced for binaural stimulation. Additionally, the sum of cortical activation volumes induced by monaural left and right stimulation was approximately 30% smaller than for binaural stimulation, indicating either inhibitory mechanisms or neuronal facilitation within the auditory pathways. For monaural deaf subjects the lateralization ratio between left to right response was just 1.3 towards the contralateral hemisphere of the healthy ear, which is comparable to binaural responses of normal-hearing subjects. This observation seems to indicate a plasticity or a reorganization of auditory pathways of monaural deaf patients.

Adult↗
PubMed1998

Ascorbic acid, a vitamin, is observed by in vivo 13C nuclear magnetic resonance spectroscopy of rat liver.

The first in vivo detection of a vitamin with nuclear magnetic resonance (NMR) is reported for mammalian liver. Vitamin C, also known as ascorbic acid, was monitored noninvasively in rat liver by "whole body" 13C NMR spectroscopy at high field after infusion of [1,2-13C2]glucose into anesthetized rats. Generally, the carbon resonances of ascorbic acid overlap with those of other highly abundant cellular metabolites, thus precluding their observation in situ. This problem was resolved by taking advantage of the 13C-13C spin couplings introduced by the two covalently bound 13C nuclei in [1,2-13C2]glucose. During glucose metabolism, [5,6-13C2]ascorbic acid was synthesized, which also exhibited characteristic 13C homonuclear spin couplings. This feature enabled the spectral discrimination of ascorbic acid from overlapping singlet resonances of other metabolites. Quantitative analysis of the spin-coupling patterns provided an estimate of the turnover rate of hepatic ascorbic acid in vivo (1.9 +/- 0.4 nmol.min-1.g-1) and a novel approach toward a better understanding of optimal ascorbic acid requirements in humans. The results obtained in vivo were confirmed with high-resolution proton and 13C NMR spectroscopy of liver extracts.

Animals↗
PubMed1997

Interaction of Alzheimer beta-amyloid peptide(1-40) with lipid membranes.

The beta-amyloid peptide beta AP(1-40), a 40-amino acid residues peptide, is one of the major components of Alzheimer's amyloid deposits. beta AP(1-40) exhibits only a limited solubility in aqueous solution and undergoes a concentration-dependent, cooperative random coil reversible beta-structure transition for Cpep > 10 microM [Terzi, E., Hölzemann, G., and Seelig, J. (1995) J. Mol. Biol. 252, 633-642]. In the presence of acidic lipid, the equilibrium is shifted further toward beta-structured aggregates. We have now characterized the lipid-peptide interaction using circular dichroism (CD) spectroscopy, lipid monolayers, and deuterium and phosphorus-31 solid-state nuclear magnetic resonance (NMR). CD spectroscopy revealed a distinct interaction between beta AP(1-40) and negatively charged unilamellar vesicles. In addition to the random coil reversible beta-structured aggregate equilibrium at low lipid-to-peptide (L/P) ratios, a beta-structure -->alpha-helix transition was observed at L/P > 55. beta AP(1-40) was found to insert into acidic monolayers provided the lateral pressure was low (20 mN/m). The extent of incorporation increased distinctly with the content of acidic lipid in the monolayer. However, at a lipid packing density equivalent to that of a bilayer (lateral pressure > or = 32 mN/m), no insertion of beta AP(1-40) was observed. The lipid molecular structure in the presence of beta AP(1-40) was studied with NMR. Phosphatidylcholine (PC) was selectively deuterated at the choline headgroup and at the cis-double bond of the oleic acyl chain and mixed with phosphatidylglycerol (PG). Phosphorus-31 NMR showed that the lipid phase retained the bilayer structure at all lipid-to-protein ratios. Deuterium NMR revealed no change in the headgroup conformation of the choline moiety or in the flexibility and ordering of the hydrocarbon chains upon the addition of beta AP-(1-40). It can be concluded that beta AP(1-40) binds electrostatically to the outer envelope of the polar headgroup region without penetrating between the polar groups. The data suggest a new mechanism of helix formation induced by the proper alignment of five positive charges of beta AP(1-40) on the negatively charged membrane template.

Amino Acid Sequence↗
PubMed1997

Inhibition of the electrostatic interaction between beta-amyloid peptide and membranes prevents beta-amyloid-induced toxicity.

The accumulation of beta-amyloid peptides (Abeta) into senile plaques is one of the hallmarks of Alzheimer disease. Aggregated Abeta is toxic to cells in culture and this has been considered to be the cause of neurodegeneration that occurs in the Alzheimer disease brain. The discovery of compounds that prevent Abeta toxicity may lead to a better understanding of the processes involved and ultimately to possible therapeutic drugs. Low nanomolar concentrations of Abeta1-42 and the toxic fragment Abeta25-35 have been demonstrated to render cells more sensitive to subsequent insults as manifested by an increased sensitivity to formazan crystals following MTT (3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) reduction. Formation of the toxic beta-sheet conformation by Abeta peptides is increased by negatively charged membranes. Here we demonstrate that phloretin and exifone, dipolar compounds that decrease the effective negative charge of membranes, prevent association of Abeta1-40 and Abeta25-35 to negatively charged lipid vesicles and Abeta induced cell toxicity. These results suggest that Abeta toxicity is mediated through a nonspecific physicochemical interaction with cell membranes.

Amyloid beta-Peptides↗
PubMed1997

Heat changes in lipid membranes under sudden osmotic stress.

Closed lipid vesicles act as osmometers increasing or decreasing their volume under the influence of osmotic gradients. The enthalpy changes accompanying membrane compression or expansion have not been measured yet, and first results obtained with high-sensitivity titration calorimetry are reported here. Phospholipid vesicles suspended in and in equilibrium with an electrolyte or nonelectrolyte with a defined initial concentration of c(i), were injected into a solution with a final concentration of c(f), and the heat changes were monitored with a titration microcalorimeter. Osmotic compression (delta c = c(f) - c(i) > 0) produced an exothermic heat change with deltaH approximately -500 +/- 100 cal/mol and osmotic expansion (delta c < 0) an endothermic heat change with deltaH approximately 1000 +/- 200 cal/mol; both results normalized to a concentration gradient of delta c = 1 M NaCl. The heats of compression and expansion varied linearly with the lipid content and the size of the osmotic gradient but were independent of the vesicle size. The cubic thermal expansion coefficient alpha(v) which equals (1/V)(deltaV/deltaT)p could be derived and was found to be 1.25 x 10(-3) and 2.5 x 10(-3) K(-1) for the compressed and expanded bilayer vesicles, respectively. The entropy changes associated with compression and expansion could be estimated. Compression of the membrane led to a negative entropy change and increased the hydrocarbon chain order. Expansion of the membrane was accompanied by a positive entropy change which can be explained, in part, by more disordered hydrocarbon chains. Vesicle expansion and compression thus appear to be asymmetric as far as the thermodynamic driving force is concerned.

Calorimetry↗
PubMed1997

Assignment of glial brain tumors in humans by in vivo 1H-magnetic resonance spectroscopy and multidimensional metabolic classification.

This study presents a simple approach for the noninvasive assignment of glial brain tumors according to malignancy by single-voxel proton magnetic resonance spectroscopy at short echo times (TE < or = 50 milliseconds). Based on peak area ratios, a five-dimensional data set was obtained for each investigated subject. This vector was then projected along metabolic coordinates in a two-dimensional metabolic space. These coordinates had been determined in a previous study (Hagberg G et al., 1995, Magn Reson Med 34: 242-252). Tumor assignment was done without any knowledge of histology by comparing the location of the new cases to the features of the previous study. All 11 investigated glioblastomas multiforme, as well as 4 of 5 astrocytomas grade II, could easily be assigned to the groups of high- and low-grade tumors, respectively. Classification was more difficult in the case of a cystic astrocytoma grade II and one astrocytoma grade III. Two spectra measured in normal-appearing matter of glioblastoma patients were not classified as healthy. Using single-voxel proton magnetic resonance spectroscopy at short echo times with the knowledge of a base study, a straightforward, fast, and noninvasive differential diagnosis of glial brain tumors is possible.

Astrocytoma↗
PubMed1997

Interaction of octyl-beta-thioglucopyranoside with lipid membranes.

Octyl-beta-thioglucopyranoside (octyl thioglucoside, OTG) is a nonionic surfactant used for the purification, reconstitution, and crystallization of membrane proteins. The thermodynamic properties of the OTG-membrane partition equilibrium are not known and have been investigated here with high-sensitivity titration calorimetry. The critical concentration for inducing the bilayer <==> micelle transition was determined as cD* = 7.3 mM by 90 degree light scattering. All thermodynamic studies were performed well below this limit. Sonified, unilamellar lipid vesicles composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) with and without cholesterol were employed in the titration calorimetry experiments, and the temperature was varied between 28 degrees C and 45 degrees C. Depending on the surfactant concentration in the membrane, the partition enthalpy was found to be exothermic or endothermic, leading to unusual titration patterns. A quantitative interpretation of all titration curves was possible with the following model: 1) The partitioning of OTG into the membrane follows a simple partition law, i.e., Xb = Kc(D,f), where Xb denotes the molar amount of detergent bound per mole of lipid and c(D,f) is the detergent concentration in bulk solution. 2) The partition enthalpy for the transfer of OTG from the aqueous phase to the membrane depends linearly on the mole fraction, R, of detergent in the membrane. All calorimetric OTG titration curves can be characterized quantitatively by using a composition-dependent partition enthalpy of the form deltaHD(R) = -0.08 + 1.7 R (kcal/mol) (at 28 degrees C). At low OTG concentrations (R < or = 0.05) the reaction enthalpy is exothermic; it becomes distinctly endothermic as more and more surfactant is incorporated into the membrane. OTG has a partition constant of 240 M(-1) and is more hydrophobic than its oxygen-containing analog, octyl-beta-D-glucopyranoside (OG). Including a third nonionic amphiphile, octa(ethyleneoxide) dodecylether (C12EO8), an empirical relation can be established between the Gibbs energies of membrane partitioning, deltaGp, and micelle formation, deltaGmic, with deltaGp = 1.398 + 0.647 deltaGmic (kcal/mol). The partition constant of OTG is practically independent of temperature and of the cholesterol content of the membrane. In contrast, the partition enthalpy shows a strong temperature dependence. The molar specific heat capacity of the transfer of OTG from the aqueous phase to the membrane is deltaCp = -98 cal/(mol x K). The OTG partition enthalpy is also dependent on the cholesterol content of the membrane. It increases by approximately 1 kcal/mol at 50 mol% cholesterol. As the partition constant remains unchanged, the increase in enthalpy is compensated for by a corresponding increase in entropy, presumably caused by a restructuring of the membrane hydration layer.

Calorimetry↗
PubMed1997

Octyl-beta-D-glucopyranoside partitioning into lipid bilayers: thermodynamics of binding and structural changes of the bilayer.

The interaction of the nonionic detergent octyl-beta-D-glucopyranoside (OG) with lipid bilayers was studied with high-sensitivity isothermal titration calorimetry (ITC) and solid-state 2H-NMR spectroscopy. The transfer of OG from the aqueous phase to lipid bilayers composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) can be investigated by employing detergent at concentrations below the critical micellar concentration; it can be defined by a surface partition equilibrium with a partition coefficient of K = 120 +/- 10 M-1, a molar binding enthalpy of delta H degrees D = 1.3 +/- 0.15 kcal/mol, and a free energy of binding of delta G degrees D = -5.2 kcal/mol. The heat of transfer is temperature dependent, with a molar heat capacity of delta CP = -75 cal K-1 mol-1. The large heat capacity and the near-zero delta H are typical for a hydrophobic binding equilibrium. The partition constant K decreased to approximately 100 M-1 for POPC membranes mixed with either negatively charged lipids or cholesterol, but was independent of membrane curvature. In contrast, a much larger variation was observed in the partition enthalpy. delta H degrees D increased by about 50% for large vesicles and by 75% for membranes containing 50 mol% cholesterol. Structural changes in the lipid bilayer were investigated with solid-state 2H-NMR. POPC was selectively deuterated at the headgroup segments and at different positions of the fatty acyl chains, and the measurement of the quadrupolar splittings provided information on the conformation and the order of the bilayer membrane. Addition of OG had almost no influence on the lipid headgroup region, even at concentrations close to bilayer disruption. In contrast, the fluctuations of fatty acyl chain segments located in the inner part of the bilayer increased strongly with increasing OG concentration. The 2H-NMR results demonstrate that the headgroup region is the most stable structural element of the lipid membrane, remaining intact until the disordering of the chains reaches a critical limit. The perturbing effect of OG is thus different from that of another nonionic detergent, octaethyleneglycol mono-n-dodecylether (C12E8), which produces a general disordering at all levels of the lipid bilayer. The OG-POPC interaction was also investigated with POPC monolayers, using a Langmuir trough. In the absence of lipid, the measurement of the Gibbs adsorption isotherm for pure OG solutions yielded an OG surface area of AS = 51 +/- 3 A2. On the other hand, the insertion area AI of OG in a POPC monolayer was determined by a monolayer expansion technique as AI = 58 +/- 10 A2. The similar area requirements with AS approximately AI indicate an almost complete insertion of OG into the lipid monolayer. The OG partition constant for a POPC monolayer at 32 mN/m was Kp approximately 320 M-1 and thus was larger than that for a POPC bilayer.

Calorimetry↗